The utility of immunohistochemistry for detecting mycobacterial infections in bronchoalveolar lavage & bronchial washings

Background & objectives: Tuberculosis, most commonly caused by Mycobacterium tuberculosis (MTB), is an infectious bacterial disease, with a major impact on global health. In this study, immunohistochemistry (IHC), acid-fast bacilli (AFB) culture and Ziehl-Neelsen (ZN) staining, techniques were compared on bronchoalveolar lavage (BAL) and bronchial washings (BW) with respect to sensitivity and specificity for detecting mycobacteria, taking culture as the gold standard. Methods: Consecutive BAL and BW specimens were included in the study, over a period of one year for which AFB cultures were available. Samples with diagnosis other than inflammatory pathology such as malignancies or inadequate samples were excluded. A total of 203 BAL and BW specimens from patients with age ranging from 14 to 86 yr were analyzed for the presence of mycobacteria. The utility and efficacy of ZN stain and IHC in detecting mycobacteria was tested using AFB culture as a gold standard. Results: Out of 203 cases, 10.3 per cent (n=21) were positive on AFB culture. Of these, 5.9 per cent (n=12) smears were positive for ZN stain, whereas IHC positivity was seen in 8.4 per cent (n=17) of the cases. ZN staining had a sensitivity of 57.1 per cent and a specificity of 100 per cent whereas, IHC had a sensitivity of 81 per cent and a specificity of 81.9 per cent. Interpretation & conclusions: Comparison with AFB culture (gold standard), IHC was found to be superior to ZN stain in terms of sensitivity, whereas ZN stain was found to be superior to IHC in terms of specificity. These findings therefore suggest that IHC may be a useful adjunct to ZN stain in the detection of mycobacteria in specimens from the respiratory tract.

Prevalence of systemic and local risk factors for pulmonary non-tuberculous mycobacterial disease in Japan: a single-institution study / Journal of Rural Medicine

Objective: To identify the prevalence of risk factors for pulmonary non-tuberculous mycobacterial (NTM) disease in a Japanese population.Patients and Methods: We reviewed 337 consecutive Japanese patients (210 women) with pulmonary NTM disease, including 225 patients with Mycobacterium avium complex (MAC) disease (95.8%) at our hospital during 2006–2017. We calculated the prevalence of risk factors reported in Western countries among mycobacterial species.Results: Pulmonary MAC disease cases comprised 78.2% of pulmonary NTM patients in their 40s, increasing to 100% at age ≥80 years. Body mass index (BMI) was <18.5 in approximately 40% of patients, which was significantly higher than the prevalence of underweight in the Japanese population. The percentage of male heavy smokers (Brinkman index ≥600) was 58.2% of pulmonary NTM disease and was high for all mycobacterial species. In pulmonary MAC disease, systemic factors were observed in the order of malignant tumors (other than lung cancer), diabetes, rheumatoid arthritis, and tuberculosis. Local factors were observed in the order of bronchiectasis, chronic obstructive pulmonary disease, lung cancer, and bronchial asthma.Conclusion: The risk factors reported in Western countries were relatively highly prevalent among Japanese pulmonary NTM disease patients. This observation may help elucidate disease onset mechanisms.

Actualización del manejo de microbacterias no tuberculosas / Update on the management of non-tuberculous mycobacteria

Desde el año 2007 se han generado guías de diagnóstico y tratamiento de micobacterias no tuberculosas (MNTB), la última de las cuales fue desarrollada en el año 2020 por ATS/ERS/ESCMID/IDSA, en ella se actualizan los criterios diagnósticos, los criterios para determinar el inicio de tratamiento y recomendaciones de esquema de antibióticos para las especies más frecuentes. En paralelo se han ido desarrollando terapias alternativas como la fagoterapia. El objetivo de la presente revisión es dar a conocer los cambios que traen estas últimas guías y actualizar algunas de las últimas novedades con respecto al manejo de las micobacterias no tuberculosas.

Since 2007, guidelines for diagnosis and treatment of non-tuberculous Mycobacteria have been generated, the latest of which was developed by ATS/ERS/ESCMID/IDSA, in which the diagnostic criteria, and the criteria for determining the initiation of treatment and antibiotic scheme recommendations for the most frequent species are updated. At the same time, alternative therapies such as phage therapy have been developed. The objective of this review is to show the changes that these latest guidelines bring and update some of the latest developments regarding the management of non-tuberculous Mycobacteria.

Nontuberculous mycobacteria in patients of a specialty hospital

ABSTRACT The incidence and clinical characteristics of NTM diseases in Brazil remain relatively unknown. The present study describes the diagnosis of NTM isolates, the clinical presentation and treatment outcomes. We analyzed NTM isolates in patients of a tertiary hospital in the Southeast region of Brazil, from January 2008 to July 2019. The ATS/IDSA criteria for diagnosis and treatment of these patients was applied. Mycobacterium kansasii were identified in 13/113 (11.5%) patients. In 59/113 (52.2%) patients who met the ATS criteria for disease, 29/59 (49.1%) received treatment, and 22/29 (75.8%) were cured. The major species identified was M. kansasii. The most frequent symptoms among the treated patients were dyspnea and cough, and the proportion of cured patients was high.

Clinical application strategies of bacteriophage in treatment of nontuberculous mycobacteriosis / 中华临床感染病杂志

Nontuberculous mycobacteria(NTM) diseases have become an important global public health problem attracting more and more attention because of its increasing morbidity. Mycobacteria show intrinsic and acquired resistance to multiple antibiotics, leading to higher difficulty and longer duration of treatment, and more uncertain prognosis than tuberculosis due to limited therapeutic measures. Bacterial phages are viruses that kill bacteria specifically, phage therapy for bacterial infection has been used for almost one century, now become a hot spot. This article reviews the biological characteristics, gene engineering of mycobacteriophages and its clinical applications; also discusses the existing problems in treatments of NTM with bacteriophages.

Pay attention to the diagnosis and treatment of Mycobacterium avium-intracellulare complex infection / 中华临床感染病杂志

Mycobacterium avium- intracellulare complex is the term of a group of slow growing nontuberculous mycobacterium related to human infections, which has received more and more attention in recent years, and become an important public health issue. This article reviews the progress on clinical diagnosis and treatment of the infections caused by Mycobacterium avium- intracellulare complex.

Strengthen the research of medication for the treatment of nontuberculous mycobacterial disease / 中华临床感染病杂志

The global trends of nontuberculous mycobacteria (NTM) infection and disease are both rising. Nontuberculous mycobacterial disease(NTMD) is a worldwide health burden associated with increasing morbidity, mortality, and economic costs. Antibiotic therapy is the mainstay of treatment for NTMD.Mycobacterial pathogens are intrinsically resistant to many available antibiotics, making treatment extremely challenging, especially in immunocompromised individuals and patients with underlying chronic lung conditions. Even with lengthy therapy and the use of a combination of antibiotics, only less than half NTMD patients can achieve clinical treatment success, so it is urgent to develop novel anti-NTM antibiotics. This article reviews the research progress on the medication of nontuberculous mycobacterial diseases.

Diagnostic value of neutrophil CD64 index in disseminated nontuberculous mycobacteria infection / 中华传染病杂志

Objective:To investigate the diagnostic value of neutrophil CD64 index (nCD64) in disseminated nontuberculous mycobacteria (NTM) infection.Methods:Thirty-six patients with NTM infection from January 2020 to June 2021 in Huashan Hospital, Fudan University were included. Patients were classified into groups of disseminated infection and focal infection according to their medical history and discharge diagnosis. The expressions of nCD64 in patients with focal infection and disseminated infection before treatment were collected and analyzed. Statistical analysis was performed using the Mann-Whitney U test, and the diagnostic value of nCD64 for disseminated NTM infection was analyzed using the receiver operator characteristic curve (ROC curve). Results:Among the 36 patients with NTM infection, 18 cases were focal infection (due to the low white blood cell count of the patient with myelodysplastic syndrome, the detection results were biased, which were excluded from the subsequent analysis) and 18 cases were disseminated infection. The expression of nCD64 in focal infection was 0.72(0.50, 1.55), and that in disseminated infection was 13.63(6.77, 32.31). The difference was statistically significant ( U=15.50, P<0.001). Using focal infection as a control, the area under the ROC curve for the operational characteristics of the subjects was 0.949 3 for disseminated NTM infection. The diagnostic cut-off value of nCD64 was 3.06, with the sensitivity and specificity of the disseminated NTM infection were 88.89% and 100.00%, respectively. Conclusions:In patients with NTM infection before effective treatment, the diagnostic cut-off value of nCD64 of 3.06 has high sensitivity and specificity, which is useful for the aided diagnosis of disseminated NTM infection.

Clinical characteristics of patients with acquired immunodeficiency syndrome complicated with nontuberculous mycobacteria disease / 中华传染病杂志

Objective:To explore the clinical characteristics of acquired immunodeficiency syndrome (AIDS) complicated with nontuberculous mycobacteria (NTM) disease.Methods:The clinical data of 190 patients with AIDS complicated with NTM disease diagnosed by Shanghai Public Health Clinical Center, Fudan University from January 1, 2019 to December 31, 2021 were analyzed retrospectively. NTM diseases were divided into disseminated NTM disease group and non-disseminated NTM disease group. The independent sample t test, Mann Whitney U test and chi-square test were used for statistical analysis. Results:The 190 patients with AIDS complicated with NTM disease included 182 males and eight females. The age was (42±13) years old, and the first hospital stay was 15(6, 26) days. Pneumocystis carinii pneumonia was the most common co-infection in 12.1%(23/190) of patients, 87 cases (45.8%) were disseminated NTM disease. The clinical symptoms of patients were common in fever (55.8%(106/190)), cough (50.0%(95/190)), and expectoration (28.9%(55/190)). The proportions of fatigue (31.0%(27/87) vs 7.8%(8/103)), poor appetite (21.8%(19/87) vs 10.7%(11/103)) in the AIDS patients with disseminated NTM disease group were higher than those in the non-disseminated NTM disease group, and the differences were statistically significant ( χ2=16.99, P<0.001 and χ2=4.42, P=0.036, respectively). There was no significant difference in the proportions of deaths between AIDS patients with disseminated NTM disease and those without disseminated NTM disease (17.2%(15/87) vs 12.6%(13/103), χ2=0.80, P=0.371). The most common NTM species was Mycobacterium avium (67.1%(49/190)), followed by Mycobacterium kansasii (15.1%(11/190)). Hemoglobin ((90.3±23.9) g/L vs (110.1±24.2) g/L), albumin ((29.7±5.5) g/L vs (34.7±5.6) g/L), CD4 + T lymphocyte count (11(5, 30)/μL vs 52(16, 96)/μL) and CD8 + T lymphocyte count ((362±320)/μL vs (496±352)/μL) in the disseminated NTM disease group were lower than those in non-disseminated NTM disease group ( t=-5.63, P<0.001; t=-6.18, P<0.001; Z=-5.90, P<0.001; and t=-2.73, P=0.007, respectively), while procalcitonin (0.24(0.10, 0.77) μg/L vs 0.10 (0.04, 0.51) μg/L) was higher than that in the non-disseminated NTM disease group ( Z=-3.09, P=0.002), with statistical significance. The most common imaging features were lung patch and strip shadow (67.4%(128/190)). Conclusions:The most common type of AIDS patients complicated with NTM disease is disseminated NTM disease, and Mycobacterium avium is the most common NTM species. The clinical manifestations (fatigue, anorexia) and laboratory tests (hemoglobin, albumin, procalcitonin, CD4 + T lymphocyte count, CD8 + T lymphocyte count) of AIDS patients with disseminated NTM disease and non-disseminated NTM disease are different, while the prognosis is not significantly different.

Escrofuloderma en paciente pediátrico / Scrofuloderma in a pediatric patient

La tuberculosis (TB) cutánea es una forma rara de tuberculosis extrapulmonar y puede tener diversas manifestaciones clínicas. La afectación cutánea puede producirse como resultado de inoculación exógena, diseminación contigua desde un foco de infección, o mediante la propagación hematógena desde un foco distante 1. Las formas multibacilares de localización cutánea siguen siendo, con mucho, las más comunes en los niños 2. La tuberculosis cutánea representa sólo el 1-2% de las formas extrapulmonares de TB. Se clasifica en varias variantes, y la escrofulodermia es una forma de tuberculosis endógena. Afecta a personas de todas las edades, sin embargo, los niños, los adolescentes y los ancianos se ven muy afectados, debido a la incapacidad inmunológica para contener la infección por micobacterias. La escrofulodermia puede presentarse de forma aislada o coexistir con formas pulmonares y diseminadas de TB. Se presenta como nódulos eritematosos que se fistulizan y descargan material caseoso y purulento 3. Los exámenes patológicos revelan abscesos, necrosis y granulomas de tipo tuberculoide (3). La correlación clínica, biológica, patológica y, a veces, la progresión con el tratamiento antibacilar son la clave del diagnóstico 2

Cutaneous tuberculosis (TB) is a rare form of extrapulmonary tuberculosis that can have diverse clinical manifestations. Cutaneous involvement may occur as a result of exogenous inoculation, contiguous dissemination from a focus of infection, or by hematogenous spread from a distant focus (1). Multibacillary forms of cutaneous localization remain by far the most common in children (2). Children and the elderly are greatly affected due to immunological inability to contain the mycobacterial infection. Scrofuloderma can occur in isolation or coexist with pulmonary and disseminated forms of TB. It presents with erythematous nodules that fistulize and discharge caseous and purulent material (3). Anatomopathological examinations reveal abscesses, necrosis and tuberculoid granulomas (3). Clinical, biological, pathological correlation and sometimes progression with antibacillary treatment are the key to diagnosis (2)

Rapid identification of Mycobacterium tuberculosis in cultures by molecular and immunological methods / Identificação rápida de Mycobacterium tuberculosis em culturas por método molecular e método imunológico

Objective: The study aimed to evaluate molecular and immunological methods and to propose a workflow using them for tuberculosis (TB) diagnosis routine. Methods: A cross-sectional retrospective study was performed, including 121 liquid cultures from a TB laboratory located in the extreme south of Brazil. All cultures were positive for Mycobacterium tuberculosis complex (MTBC) by in-house Polymerase Chain Reaction (PCR) using DNA extracted by the CTAB method (PCR-CTAB) for IS6110 detection. These cultures were subjected to faster tests than this one, the immunological MPT64 assay and the PCR using DNA extracted by thermal lysis method (PCR-TL), and these were evaluated for MTBC identification using PCR-CTAB as a reference method. Results: The sensitivity of MPT64 assay and PCR-TL to identify MTBC in positive cultures by PCR-CTAB were 73.6% (89/121) and 98.3% (119/121), respectively. We proposed a workflow based on the use of MPT64 assay in liquid cultures suggestive of MTBC, and in case of a negative result, we suggest the performance of PCR-TL. The PCR-CTAB is suggested only if faster tests are negative. Conclusions: Methods capable of confirming MTBC in cultures should continue to be standardized, tested, and optimized to meet the ideal requirements of simplicity, quickness, and effectiveness. The molecular and immunological methods evaluated have differences in the execution and detection of MTBC in cultures, but they are rapid tools for laboratory TB diagnosi

Objetivos: O estudo objetivou avaliar métodos molecular e imunológico e propor um fluxo de trabalho utilizando-os para a rotina de diagnóstico da tuberculose (TB). Métodos: Foi realizado um estudo transversal retrospectivo, incluindo 121 culturas líquidas de um laboratório de TB localizado no extremo sul do Brasil. Todas as culturas foram positivas para o complexo Mycobacterium tuberculosis (CMTB) por Reação em Cadeia da Polimerase (PCR) in-house para detecção do IS6110, usando DNA extraído pelo método CTAB (PCR-CTAB). Essas culturas foram submetidas a testes mais rápidos que este, o ensaio imunológico MPT64 e a PCR com DNA extraído pelo método de lise térmica (PCR-LT), e estas foram avaliadas para identificação de CMTB usando PCR-CTAB como método de referência. Resultados: A sensibilidade do ensaio MPT64 e da PCR-LT para identificar o CMTB em culturas positivas pela PCRCTAB foi de 73,6% (89/121) e 98,3% (119/121), respectivamente. Propusemos um fluxo de trabalho baseado no uso do ensaio MPT64 em culturas líquidas sugestivas de CMTB e, em caso de resultado negativo, sugerimos a realização de PCR-LT. Sugere-se a PCR-CTAB apenas se os testes mais rápidos forem negativos. Conclusões: Os métodos capazes de confirmar o CMTB em culturas devem continuar sendo padronizados, testados e otimizados para atender aos requisitos ideais de simplicidade, rapidez e eficácia. Os métodos molecular e imunológico avaliados apresentam diferenças na execução e detecção do CMTB em culturas, mas são ferramentas rápidas para o diagnóstico laboratorial da TB.

Micobacteriosis atípica post mesoterapia / Atypical mycobacteriosis post mesotherapy

Resumen La utilización de procedimientos estéticos que mejoren la imagen corporal está en constante crecimiento, y también las infecciones asociadas a ellos, como las micobacteriosis atípicas. La meso terapia es un procedimiento mínimamente invasivo que consiste en la aplicación de sustancias que buscan estimular la dermis y el tejido celular subcutáneo, para el tratamiento de la celulitis y el rejuvenecimiento de la piel. Reportamos un caso de infección micobacteriana posterior a una mesoterapia en glúteos y muslos que se presentó como abscesos subcutáneos, que respondieron satisfactoriamente al tratamiento antibiótico prolonga do con claritromicina y trimetoprima-sulfametoxazol. Se han informado infecciones asociadas a mesoterapia en España, América Latina y el Caribe, que tendrían posiblemente un origen común: la falta de controles sanitarios. Destacamos la importancia de estar alertados sobre estas complicaciones infecciosas y la necesidad de reforzar las medidas de seguridad necesarias para evitarlas.

Abstract The use of aesthetic procedures that improve body image is constantly growing, as well as infections associated with them, such as atypical mycobacteriosis. Mesotherapy is a minimally invasive aesthetic procedure that consists of the application of substances that seek to stimulate the dermis and subcutaneous cellular tissue, for the treatment of cellulite and skin rejuvenation. We report a case of mycobacterial infection after mesotherapy in the buttocks and thighs that appeared as subcutaneous abscesses, they responded satisfactorily to prolonged antibiotic treatment with clarithromycin and trimethoprim-sulfamethoxazole. Infections associated with mesotherapy have been reported in Spain, Latin America and the Caribbean, all possibly related to lack of health controls. We emphasize the importance of being aware of these infectious complications and the need to reinforce the necessary security measures to avoid them.

Levantamento epidemiológico dos casos de tuberculose no Brasil e ações alternativas para auxiliar no tratamento / Epidemiological survey of tuberculosis cases in Brazil and alternative actions to aid in treatment / Estudio epidemiológico de los casos de tuberculosis en Brasil y acciones alternativas para ayudar al tratamiento

tuberculose é uma das doenças infectocontagiosas de maior importância no Brasil e no mundo. Afeta de forma importante populações em situação de vulnerabilidade social e econômica. O objetivo deste estudo foi realizar um levantamento do número de casos notificados de tuberculose no Brasil nos últimos 10 anos (2011 a 2021), avaliar os fatores que afetam a transmissão, bem como discutir o tratamento padrão e com fitoterápicos. O levantamento epidemiológico dos casos de tuberculose no Brasil de janeiro de 2011 a dezembro de 2021 foi realizado dentre os notificados pelo Sistema de Informação de Agravos de Notificação (SINAN). Os resultados indicaram um aumento linear de casos a partir de 2017 com 90.776 casos diagnosticados, em 2018 (94.720) e 2019 (96.655). Acredita-se que o aumento linear da tuberculose neste período pode estar relacionado principalmente com o aumento da pobreza, contudo o compartilhamento de utensílios durante o uso de narguilé podem representar fatores de risco para tuberculose. Seis plantas medicinais afetam diretamente as micobactérias (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata) e cinco plantas com atividade antibacteriana auxiliam no trato respiratório (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). Contudo, a tuberculose é uma doença reemergente sendo necessária a adoção de políticas públicas que intensifiquem e implementem medidas sócio-educativas para a implantação do uso de fitoterápicos como medida complementar.

Tuberculosis is one of the most important infectious diseases in Brazil and worldwide. It significantly affects populations in situations of social and economic vulnerability. This study aimed to survey the number of reported tuberculosis cases in Brazil in the last 10 years (2011 to 2021) to assess the factors that affect the transmission and discuss standard and herbal treatments. The epidemiological survey of tuberculosis cases in Brazil from January 2011 to December 2021 was carried out among those notified by the Notifiable Diseases Information System (SINAN). The results indicated a linear increase in cases from 2017, with 90,776 diagnosed cases, in 2018 (94,720) and 2019 (96,655). It is believed that the linear increase in tuberculosis in this period may be mainly related to the increase in poverty. However, the sharing of utensils during the use of hookah may represent risk factors for tuberculosis. Six medicinal plants directly affect mycobacteria (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata), and five plants with antibacterial activity help in the respiratory tract (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). However, tuberculosis is a re-emerging disease, and it is necessary to adopt public policies that intensify and implement socio-educational measures for using herbal medicines as a complementary measure.

La tuberculosis es una de las enfermedades infecciosas más importantes en Brasil y en el mundo. Afecta significativamente a las poblaciones en situación de vulnerabilidad social y económica. El objetivo de este estudio fue realizar una encuesta sobre el número de casos notificados de tuberculosis en Brasil en los últimos 10 años (2011 a 2021), para evaluar los factores que afectan a la transmisión, así como para discutir el tratamiento estándar y con fitoterapias. La encuesta epidemiológica de los casos de tuberculosis en Brasil desde enero de 2011 hasta diciembre de 2021 se realizó entre los notificados por el Sistema de Informação de Agravos de Notificação (SINAN). Los resultados indicaron un aumento lineal de casos desde 2017 con 90.776 casos diagnosticados, en 2018 (94.720) y 2019 (96.655). Se cree que el aumento lineal de la tuberculosis en este periodo puede estar relacionado principalmente con el aumento de la pobreza, aunque el hecho de compartir los utensilios durante el uso de la shisha puede representar factores de riesgo para la tuberculosis. Seis plantas medicinales afectan directamente a las micobacterias (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata) y cinco plantas con actividad antibacteriana ayudan a las vías respiratorias (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). Sin embargo, la tuberculosis es una enfermedad reemergente siendo necesaria la adopción de políticas públicas que intensifiquen e implementen medidas socioeducativas para la implementación del uso de fitoterápicos como medida complementaria.

Eventos de precipitación extrema aumentan la circulación de patógenos infecciosos: detección de rotavirus, norovirus, astrovirus, adenovirus entérico y micobacterias no tuberculosas en agua de las inundaciones en Asunción, Paraguay, año 2014

Las precipitaciones extremas representan uno de los eventos naturales climáticos más importantes y pueden originar inundaciones devastadoras. De junio a agosto del 2014 se registró una de las más graves inundaciones en la historia de la ciudad de Asunción. Ocasionó un incremento considerable del nivel del río Paraguay y el desplazamiento de 300.000 personas a campamentos provisionales. Debido a que el contacto directo con el agua de inundación, el consumo de agua contaminada y la congregación de los afectados en refugios provisorios son factores de riesgo para enfermedades infecciosas, el objetivo de este estudio fue la implementación de una metodología estandarizada para la concentración y detección de virus entéricos y micobacterias no tuberculosas, por PCR en tiempo real y PCR-asociada al análisis de restricción enzimática (PRA), en muestras de agua de inundaciones y el reporte de los patógenos detectados en las zonas afectadas de Asunción y en la Bahía del Río Paraguay. La metodología propuesta demostró poseer buena sensibilidad y se registró la presencia de rotavirus, norovirus (genogrupos I y II), astrovirus, adenovirus entéricos y micobacterias no tuberculosas en 50% (N=4/8) de las muestras de los barrios Sajonia, San Jerónimo y Ricardo Brugada, Chacarita. Además, reportamos datos secundarios de casos de enfermedades infecciosas, registrados en los servicios de salud de los barrios afectados durante el periodo de inundación.

Extreme rainfall represents one of the most important natural climatic events and can cause devastating floods. From June to August 2014, one of the most serious floods in the history of the city of Asunción was recorded. It caused a considerable increase in the level of the Paraguay River and the displacement of 300,000 people to temporary camps. Since direct contact with flood water, consumption of contaminated water and the congregation of those affected in temporary shelters are risk factors for infectious diseases, the objective of this study was the implementation of a standardized methodology for the concentration and detection of enteric viruses and non-tuberculous mycobacteria, by real-time PCR and PCR-associated enzyme restriction analysis (PRA), in samples of flood water and the report of the pathogens detected in the affected areas of Asunción and in the Bay of the Paraguay River. The proposed methodology proved to have good sensitivity and the presence of rotavirus, norovirus (genogroups I and II), astrovirus, enteric adenovirus and non-tuberculous mycobacteria was recorded in 50% (N=4/8) of the samples from the Sajonia, San Jeronimo and Ricardo Brugada, Chacarita neighborhoods. In addition, we report secondary data on cases of infectious diseases, registered in the health services of the affected neighborhoods during the flood period.

Contribución del uso de XPERT MTB/RIF y su costo-efectividad en el diagnóstico de tuberculosis pulmonar y la resistencia a rifampicina: una comparación con métodos diagnósticos no moleculares / Contribution of the use of xpert MTB/RIF and its cost-effectiveness in the diagnosis of pulmonary tuberculosis and resistance to rifampicin: a comparison with non molecular diagnostic methods

Resumen Introducción: La tuberculosis es un problema de salud pública; su control requiere diagnóstico temprano y tratamiento oportuno. Xpert MTB/RIF® es una tecno logía diagnóstica basada en PCR en tiempo real, detecta el Complejo Mycobacterium tuberculosis y la susceptibilidad a rifampicina. Objetivo: Determinar la contribución del Xpert MTB/RIF y su costo-efectividad en la detección de tuberculosis y la resistencia a rifampicina en muestras respirato rias al compararlo con métodos de diagnóstico no moleculares Materiales y Métodos: Se analizaron 1.574 muestras de pacientes con sospecha de tuberculosis pulmonar que fueron procesadas para microscopía con coloración fluorescente de auramina-rodamina, Xpert MTB/RIF y cultivo en BACTEC MGIT 960. Los resultados obtenidos se compararon entre los métodos no moleculares y los moleculares para la detección de M. tuberculosis y susceptibilidad a rifampicina y se realizó un análisis comparativo de costos y costo efectividad. Resultados: 19,2% de las muestras fueron positivas por alguna de las técnicas usadas. Xpert MTB/RIF detectó M. tuberculosis en 90,4% del total de muestras positivas con un índice Kappa de 0,77 (IC95%: 0,74-0,82) comparado con el cultivo. La resistencia a rifampicina por Xpert fue 8,1%, sensibilidad 94,1% (IC95%: 73,0-99,0%), especificidad 98,4% (IC95%: 95,5-99,5%) y Kappa de 0,88 (IC95%: 0,76-1,00). La razón incremental de costo efectividad (RICE) fue menor en Xpert MTB/RIF comparada con el cultivo. Conclusión: Xpert MTB/RIF es una prueba eficiente y costo efectiva en la detección de casos de M. tuberculosis en muestras pulmonares comparado con los mé todos de diagnóstico basados en cultivo, sin embargo y a diferencia del Xpert MTB/RIF, estos pueden aportar en el diagnóstico con el aislamiento de especies de micobacterias no tuberculosas y la susceptibilidad a isoniazida y otros medicamentos.

Abstract Introduction: Tuberculosis is a public health problem its control requires early diagnosis and timely treatment. Xpert MTB/RIF is a real-time PCR based diagnostic technology, detects the Mycobacterium tuberculosis complex and rifampicin resistance. Objective: To determine the contribution of Xpert MTB/RIF and its cost-effectiveness in the detection of potential positive cases for tuberculosis and resistance to rifampicin in respiratory samples comparatively with diagnostic non molecular methods Materials and Methods: From 2013 to 2015, 1.574 clinical samples of patients with suspected pulmonary tuberculosis were evaluated by smear microscopy using auramina-rodamina stain, Xpert and culture in liquid medium BACTEC MGIT 960®. Results: 19,2% of the samples were positive for any of the methods used, Xpert detected M. tuberculosis in 90,4% of the positive samples and the concordance between Xpert and cultures had a Kappa index of 0,71 (IC95%: 0,62-0,72). Xpert identified resistance to rifampicin in 8,1% of the clinical samples studied with a sensitivity 94.1% (IC95%: 73,0-99,0%), specificity 98,4% (IC95%: 95,5-99,5%) and Kappa index 0,88 (IC95%: 0,76-1,00). Xpert had an incremental cost effectiveness ratio lower than culture (RICE). Conclusion: Xpert MTB/Rif is efficient diagnostic technique and comparable with culture in cost effectiveness for pulmonary tuberculosis diagnosis. However, culture based methods, in contrast to Xpert, may allow the isolation and identification of non tuberculosis mycobacterial species and the possibility to perform susceptibility for other antituberculous drugs.

Queratitis por Mycobaterium abscessus: comunicación del primer caso en Paraguay / Keratitis due to Mycobaterium abscessus: first case report in Paraguay

Resumen Se presenta un caso clínico de queratitis por Mycobacterium abscessus en una mujer de 76 años, residente en la ciudad de Asunción, sin traumatismo ni cirugía ocular previa y con antecedente de una queratouveitis herpética. Por tratarse de una queratitis causada por un agente etiológico poco frecuente y por la importancia de un diagnóstico correcto y oportuno para la instauración del tratamiento adecuado, se comunica el primer caso de queratitis por micobacterias en Paraguay.

Abstract We present a clinical case of keratitis caused by M. abscessus in a 76-year-old female patient, resident in the city of Asunción, without trauma or previous ocular surgery and with a history of herpetic keratouveitis. Because it is a keratitis caused by a rare etiological agent and because of the importance of a correct and timely diagnosis for the establishment of appropriate treatment, the present case is reported, the first of Mycobacteria keratitis in Paraguay.

Clinical and laboratory characteristics of 71 patients with acquired immunodeficiency syndrome complicated with disseminated nontuberculous mycobacterial disease / 中华传染病杂志

Objective:To explore the clinical and laboratory characteristics of acquired immunodeficiency syndrome (AIDS) patients complicated with disseminated nontuberculous mycobacterial (NTM) disease, in order to provide basis for clinical therapy.Methods:The clinical findings, imaging and etiological data of the 71 AIDS patients complicated with disseminated NTM disease admitted to Beijing You′an Hospital from June 2016 to June 2021 were retrospectively analyzed.Results:Among 71 patients with disseminated NTM disease, the most common initial symptom was fever, followed with cough, expectoration, fatigue, poor appetite, abdominal pain and diarrhea. Seventy point four percent (50/71) of these patients had at least two comorbidities, with oral candida infection, cytomegalovirus infection, syphilis, pneumocystis pneumonia and bacterial pneumonia being the most common. Hemoglobin ((87.8±24.2) g/L) and albumin ((27.3±7.0) g/L) levels significantly decreased, while erythrocyte sedimentation rate ((59.8±28.6) mm/1 h) and C-reactive protein ((74.7±50.8) mg/L) levels increased in most cases. The median CD4 + T lymphocyte count was 7×10 6/L. The median time of positive blood culture of NTM was 260 h. Among the 71 patients, 40 cases (56.3%) were infected with Mycobacterium avium, 15 cases (21.1%) with Mycobacterium intracellulare, 10 cases (14.1%) with Mycobacterium colombiense, three cases (4.2%) with Mycobacterium marseillense and three cases (4.2%) with Mycobacterium kansasii.The frequent imaging findings were patchy and nodular shadows in lungs, and most patients had mediastinal or hilar lymph node enlargement and splenomegaly. Conclusions:AIDS complicated with disseminated NTM disease is prevalently occurred in patients with severe immune deficiency, and most of the bacteria belong to the Mycobacterium avium- intracellulare complex. Early obtaining positive etiological results of NTM is essential to guide the correct clinical diagnosis and accurate treatment.

Clinical application of gene chip technology in diagnosis and treatment of silicosis complicated with mycobacterial infection / 环境与职业医学

Background Gene chip technology has been increasingly used in the diagnosis and treatment of common tuberculosis. However, its role in the diagnosis and treatment of silicosis complicated with mycobacterial infection remains unclear. Objective To evaluate the application value of gene chip technology in the diagnosis and treatment of silicosis complicated with mycobacterial infection. Methods From January 2019 to June 2021, 197 silicosis patients suspected to be complicated with mycobacterial infection in Quanzhou First Hospital Affiliated to Fujian Medical University were enrolled in this study. The etiology evaluation for the 197 patients was conducted by acid-fast staining of sputum smear (sputum smear method), culture of Mycobacterium tuberculosis of sputum (sputum culture method), and gene chip technology of bronchoalveolar lavage fluid (BALF); and for 80 patients among them, acid-fast staining of BALF (BALF smear method) and culture of Mycobacterium tuberculosis of BALF (BALF culture method) were additionally performed. The positive rates and consistency were assessed using intraclass correlation coefficient (ICC). Test for Mycobacterium tuberculosis drug resistance mutation gene was added for patients with Mycobacterium tuberculosis complex by BALF gene chip technology. Results The average age of the 197 patients was (53.1±9.1) years, and the average dust exposure time was (21.1±9.4) years, including 192 males and 5 females. There were 8 cases with stage I silicosis, 17 cases with stage II silicosis, and 172 cases with stage III silicosis. Among them, 11.2% were positive for sputum smear; 24.4% were positive for sputum culture, and 36.0% were positive by gene chip of BALF. The difference between the three methods was statistically significant (P<0.05). The result of consistency test for the three methods showed that the ICC was 0.539 (P<0.001). Among the 80 patients, there was a significant difference in the positive rates of the five methods (χ2=25.23, P<0.001). The results of Bonferroni test showed statistically significant pair-wise differences between BALF culture method and sputum smear method, BALF culture method and BALF smear method, BALF gene chip method and sputum smear method, BALF gene chip method and BALF smear method (P<0.05), while there were no statistically significant differences between the other pairs (P>0.05). The result of consistency test for the five methods showed that the ICC was 0.586 (P<0.001). Among the 71 BALF gene chip positive cases, 59 cases reported positive Mycobacterium tuberculosis complex (17 cases were positive in the first-line anti-tuberculosis resistance test, and 2 cases were found positive quinolone resistance gene in the second-line anti-tuberculosis resistance test), and received regular anti-tuberculosis treatment, among them 45 cases improved and 14 cases were stable; 12 cases reported non-tuberculous mycobacteria cases, among them 5 cases received anti-non-tuberculous mycobacteria treatment (4 cases improved and 1 case was stable), and 7 cases with mild symptoms did not receive anti-non-tuberculous mycobacteria treatment. Conclusion Compared with sputum smear, sputum culture, and other traditional methods, gene chip technology of BALF can improve the positive rate of pathogenic diagnosis of silicosis complicated with mycobacterial infection, and can also quickly identify whether it is non-tuberculous mycobacteria infection or drug-resistant Mycobacterium tuberculosis infection, which is helpful to adjust treatment as soon as possible.

Detection of non-tuberculosus mycobacteria (NTMs) in lung samples using 16S rRNA

BACKGROUND Non-tuberculous mycobacteria (NTMs) cause diseases known as mycobacteriosis and are an important cause of morbidity and mortality. The diagnosis of pulmonary disease caused by NTM is hampered by its clinical similarity with tuberculosis (TB) and by the lack of an accurate and rapid laboratory diagnosis. OBJECTIVES Detect DNA from NTMs directly from lung samples using real-time polymerase chain reaction (qPCR) for amplification of 16S rRNA. Additionally, DNA sequencing (hsp65 and rpoB genes) was used to identify the species of MNTs. METHODS A total of 68 sputum samples (54 with suspected NTMs and 14 with TB) from patients treated at a referral hospital were used. FINDINGS Of these, 27/54 (50%) were qPCR positive for NTMs and 14/14 TB patients (controls) were qPCR negative with an almost perfect concordance (Kappa of 0.93) with the Mycobacterium spp. culture. Sequencing confirmed the presence of NTM in all positive samples. The most common species was Mycobacterium gordonae (33%), followed by Mycobacterium abscessus (26%), Mycobacterium fortuitum (22%), Mycobacterium avium (15%) and Mycobacterium peregrinum (4%). MAIN CONCLUSIONS The qPCR technique for detecting NTMs targeting 16S rRNA has the potential to detect NTMs and rapidly differentiate from Mycobacterium tuberculosis. However, it is necessary to identify the species to help in the differential diagnosis between disease and contamination, and to guide the choice of the therapeutic scheme.

Epidemiological profile and follow-up of patients with pulmonary disease by non-tuberculous mycobacteria in Baixada Santista area, São Paulo

ABSTRACT Objective To present the frequency and species diversity of non-tuberculous mycobacteria, estimate the prevalence of non-tuberculous mycobacterial pulmonary disease, describe the epidemiological profile, and determine the follow-up of patients with non-tuberculous mycobacterial pulmonary disease living in a region with a high burden of tuberculosis. Methods This a retrospective cohort observational study using data records obtained from the Instituto Adolfo Lutz - Santos and from the São Paulo Sistema de Vigilância de Tuberculose do Estado de São Paulo in the period between 2000 and 2009. The studied variables were: socio-demographic characteristics, current and past history of tuberculosis, aspects related to diagnosis, and treatment and associated diseases. Results We included 319 non-tuberculous mycobacteria isolates in the study, corresponding to 257 patients. The species Mycobacterium kansasii (28.5%) and Mycobacterium fortuitum (16.6%) presented the higher occurrence. In 10.9% (24) of the patients, there was a criterion for confirming a case of pulmonary disease due to non-tuberculous mycobacteria. In relation to gender and age, male and individuals over 50 years old were the most frequent. Considering the confirmed cases, 47.8% had a past history of tuberculosis. Conclusion The lack of information about the cases is evident, since pulmonary disease due to non-tuberculous mycobacteria is not mandatory. The therapeutic regimen according to the identified species is fundamental for success in combating the infections caused by non-tuberculous mycobacteria. Besides that, information about the regional epidemiology of pulmonary disease caused by non-tuberculous mycobacteria and the search for associations with other comorbidities are important to establish the correct treatment. In order to improve surveillance of pulmonary diseases by non-tuberculous mycobacteria, we suggest the implantation of a sentinel surveillance and of population-based studies.